Provided herein, inter alia, are methods and compositions for engineering T cells. The methods include contacting a T cell with a nucleic acid and one or more cyclic GMP-AMP synthase (cGAS)-stimulator of interferon gene (STING) pathway inhibitors. The methods provided herein are contemplated to increase cell viability, expansion and gene editing efficiency, thereby allowing an increase in the total number of engineered T cells.

Type V Cas proteins, for example Type V Cas proteins referred to as ZWGD, ZJHK, ZIKV, ZZFT, YYAN, ZZGY, ZKBG, ZZKD, ZXPB, ZPPX, ZXHQ, ZQKH, ZRGM, ZTAE, ZSQQ, ZSYN, ZRBH, ZWPU, ZZQE, and ZRXE Type V Cas proteins; gRNAs for Type V Cas proteins; systems comprising Type V Cas proteins and gRNAs; nucleic acids encoding the Type V Cas proteins, gRNAs and systems; particles comprising the foregoing; pharmaceutical compositions of the foregoing; and uses of the foregoing, for example to alter the genomic DNA of a cell.

Provided herein are prime editing methods and compositions for treatment of genetic disorders such as Fuchs endothelial corneal dystrophy type 3.

Disclosed herein are Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) systems for use as a medicament, the CRISPR system comprising a guide RNA (gRNA) comprising the sequence set forth in any one of SEQ ID NO: 3-74, and a CRISPR-associated endonuclease, the gRNAs targeting the Neh4 and/or Neh5 domain of NRF2. Also disclosed herein are methods of using the aforementioned gRNAs, DNA sequences encoding such gRNAs, and vectors and pharmaceutical compositions comprising such gRNAs.

The present disclosure provides improved CRISPR-Cas proteins (e.g., improved thermostability).

This invention relates generally to polymeric delivery systems. The present polymeric delivery systems may be complexed with biological agents, including nucleic acids, peptides, proteins, or small molecules, for delivery to cells. In particular, the present polymeric delivery systems may be used in gene editing.

The present invention relates to chemically inducible split protein actuators (CISPA), which utilize ligand-binding proteins or protein domains originating from humans or other organisms, which are rationally split into two fragments that reassemble only in the presence of a cognate ligand. In particular, the invention relates to their design, manufacture, structure, and uses. The designed CISPAs can be used to regulate cellular processes such as gene expression, conditionally reconstitute of the function of a protein such as enzyme activity, as biological sensors, or for other applications.

Disclosed herein are compositions and methods comprising epigenetic editing systems for epigenetic editing or cells, nucleic acids, and vectors comprising the same. Also disclosed are epigenetically modified chromosomes.

The present disclosure provides systems and methods for high throughput genetic manipulation. Particularly, systems and methods are provided for scalable gene insertions in mammalian cells, the systems and methods comprise a donor nucleic acid comprising a cargo sequence encoding one or more selectable markers; a first guide RNA complementary to at least a portion of the donor nucleic acid; a plurality of second guide RNAs each of which is complementary to at least a portion of one of a plurality of target nucleic acids; a first RNA-guided endonuclease configured to bind to the first guide RNA; a second RNA-guided endonuclease configured to bind to the plurality of second guide RNAs; or one or more nucleic acids encoding thereof.

A complex, comprising a first fusion and a second fusion, wherein one of the first fusion and the second fusion comprises a DNA methylation domain and at least one recruitment domain A, and the other fusion comprises a transcriptional repressor domain and at least one recruitment domain A; and the recruitment domain A and the recruitment domain A interact with each other, so that one of the first fusion and the second fusion or a part thereof is recruited to the vicinity of the other fusion. Also provided are use of the complex in the preparation of a product for inhibiting target gene expression and the preparation of a drug.